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LIPID BIOSYNTHESIS

Fatty acid biosynthesis-basic fundamentals Fatty acid biosynthesis-elongation and desaturation Triacylglycerols Phospholipids Cholesterol Cholesterol metabolism

Fatty Acid Biosynthesis


Synthesis Beta Oxidation

Cytosol Requires NADPH Acyl carrier protein D-isomer CO2 activation Keto saturated

Mitochondria NADH, FADH2 CoA L-isomer No CO2 Saturated keto

Rule:
Fatty acid biosynthesis is a stepwise assembly of acetyl-CoA units (mostly as malonyl-CoA) ending with palmitate (C16 saturated)

3 Phases
Activation

Elongation
Termination

ACTIVATION
O

Cofactor
Biotin
NH C 2C 2C 2C 2C H H H H O S N H C 2C 2C 2 E ZY E HC 2 H H H N M HN

CH3C~SCoA O

ATP
ADP + Pi
-OOC-CH

HCO3-

Biocytin
2C~SCoA

LY S

O
O

O C N

O NH

CO2

active carbon
S

C 2C 2C 2C 2C H H H H O

Acetyl-CoA carboxylase

Carboxybiocytin

Acetyl-CoA Carboxylase
The rate-controlling enzyme of FA synthesis
In Bacteria -3 proteins (1) Carrier protein with Biotin (2) Biotin carboxylase (3) Transcarboxylase In Eukaryotes - 1 protein (1) Single protein, 2 identical polypeptide chains (2) Each chain Mwt = 230,000 (230 kDa) (3) Dimer inactive (4) Activated by citrate which forms filamentous form of protein that can be seen in the electron microscope

Yeast Fatty Acid Synthase Complex 2,500 kDa Multienzyme Complex


6 molecules of 2 peptide chains called A and B

6 6)

A: (185,000) Acyl Carrier protein -ketoacyl-ACP synthase (condensing enzyme) -ketoacyl-ACP reductase
B: (175,000) -hydroxy-ACP dehydrase enoyl-ACP reductase palmitoyl thioesterase Fatty Acid Synthase Complex

Acyl Carrier Protein


Phosphopantetheine O HS-CH2-CH2-N-C-CH2-CH2-N-C-C-C-CH2-O-P-O-CH2-SerO OH H O Cysteamine
H

H HO CH3

ACP

Acyl carrier protein 10 kDa


O O HS-CH2-CH2-N-C-CH2-CH2-N-C-C-C-CH2-O-P-O-P-O-CH2 O O OH H O O O O-P-O OH
H

H HO CH3

Adenine
H

Coenzyme A

OH

Initiation

Overall Reaction
Malonyl-CoA + ACP
-OOC-CH 2C~S-

CH3C~SCoA O

ACP

+ HS-CoA

CO2

HS-CoA
ACP

Acyl Carrier Protein

CH3C-CH2C~SO O

NOTE: Malonyl-CoA carbons become new COOH end Nascent chain remains tethered to ACP CO2, HS-CoA are released at each condensation

-Carbon
CH3C-CH2C~SACP

Elongation
O
O Reduction
-Ketoacyl-ACP reductase

NADPH
D isomer

H CH3C-CH2C~SHO O H CH3C- C- C~S= H O

ACP Dehydration
-Hydroxyacyl-ACP dehydrase

-H2O NADPH

ACP Reduction
Enoyl-ACP reductase

CH3CH2CH2C~S- ACP O

TERMINATION
-KS

Ketoacyl ACP Synthase

Transfer to Malonyl-CoA

Transfer to KS -S-ACP

-CH2CH2CH2C~S- ACP O

Split out CO2


CO2

Free to bind Malonyl-CoA

When C16 stage is reached, instead of transferring to KS, the transfer is to H2O and the fatty acid is released

Fatty Acid Synthase


-Ketoacyl -ACP synthase
KS

O S-C-CH2-CH2-CH3
KS

Acetyl-CoA HS CoA-SH

Acetyl-CoAACP transacylase

O CH3-CH2-CH2-C-S Enoyl-ACP reductase NADPH H+ O CH3-CH=CH-C-S -Hydroxyacyl-ACP H2O


dehydrase

NADP+

O S -C-CH3
KS -SH
KS

Initiation or priming

ACP

O S-C-CH3

SH Malonyl-CoA
Malonyl-CoACoA-SH ACP transacylase

OH

CH3-CH -CH2-C-S NADP+ NADPH H+

S
C=O CH2 C=O CH3 CO2

O S -C-CH2-COO-Keto-ACP synthase (condensing enzyme)


KS -SH

-Ketoacyl -ACP reductase

Elongation

Substrate Entry AT MT

Reduction

Thioesterase palmitate release

CE
CH2 HS Translocation HS

DH KR ER ACP

TE
SH CH2 CE

SH Translocation

TE

ACP ER KR DH

MT

AT

Thioesterase palmitate release

Reduction

Substrate Entry

Overall Reactions
Acetyl-CoA + 7 malonyl-CoA + 14NADPH + 14H+ 7H Palmitate + 7CO2 + 14NADP+ + 8 HSCoA + 6H2O

7 Acetyl-CoA + 7CO2 + 7ATP 7 malonyl-CoA +7ADP + 7Pi + 7H+

8 Acetyl-CoA + 14NADPH + 7H+ + 7ATP Palmitate + 14NADP+ + 8 HSCoA + 6H2O + 7ADP + 7Pi

PROBLEM: Fatty acid biosynthesis takes place in the cytosol. Acetyl-CoA is mainly in the Mitochondria
acetyl-CoA

How is acetyl-CoA made available to the cytosolic fatty acyl synthase?


SOLUTION: Acetyl-CoA is delivered to cytosol from the mitochondria as CITRATE

CH2COO HO-C-COO

HS-CoA

Acetyl-CoA

mitochondria
CH2COO HO-C-COO CH2COO Acetyl-CoA

CH2COO

Citrate lyase
COO C=O OAA Malate CH2 dehydrogenase COO
NADH

L-malate CO2

OAA
CO2 Pyr

COO HO-C-H L-malate CH2 COO Malic enzyme


NADP+

NADPH + H+ COO C=O Pyruvate CH3

Cytosol

Post-Synthesis Modifications
C16 satd fatty acid (Palmitate) is the product

Elongation Unsaturation Incorporation into triacylglycerols Incorporation into acylglycerol phosphates

Elongation of Chain (two systems)


R-CH2CH2CH2C~SCoA Malonyl-CoA* O (cytosol) HS-CoA OOC-CH2C~SCoA CH3C~SCoA O O CO2 Acetyl-CoA (mitochondria) R-CH2CH2CH2CCH2C~SCoA O O 1 NADPH Elongation systems are NADH found in smooth ER and 2 - H2O
3 NADPH

mitochondria

R-CH2CH2CH2CH2CH2C~SCoA O

Desaturation
Rules:
The fatty acid desaturation system is in the smooth membranes of the endoplasmic reticulum There are 4 fatty acyl desaturase enzymes in mammals designated 9 , 6, 5, and 4 fatty acyl-CoA desaturase Mammals cannot incorporate a double bond beyond 9; plants can. Mammals can synthesize long chain unsaturated fatty acids using desaturation and elongation

Rule:

The Desaturase System requires O2 and resembles an electron transport system Cyt b5 reductase
(FAD)

NADH
2

Cyt b5
3

O2

Saturated FA-CoA
1 2

NOTE: 1. System is in ER membrane

2. Both NADH and the fatty acid contribute electrons 3. Fatty acyl desaturase is considered a mixed function oxidase

Fatty acid desaturation system


C18-stearoly-CoA C18 9-oleyl-CoA + O2 + 2H+ + 2H2O Desaturase

Desaturase
2 cyt b5 Fe2+ 2 cyt b5 Fe2+ Cyt b5 2H+ + cyt b5 reductase FAD cyt b5 reductase FADH2

Cyt b5 reductase

NADH + H+

NAD+

Desaturase
Palmitoleate 16:1( 9)

Palmitate 16:0

Elongase
Stearate 18:0

Desaturase Desaturase

Essential fatty acid

Oleate 18:1( 9)
Linoleate 18:2( 9,12)

Permitted transitions in mammals

Desaturase
-Linolenate 18:3( 9,12,15) Other lipids

Desaturase Elongase

-Linolenate 18:3( 6,9,12)

Desaturase

Eicosatrienoate 20:3( 8,11,14)


Arachidonate 20:4( 5,8,11,14)